Embodiments of the invention relate to linear spectral unmixing and the reduction of artifacts in samples such as multiply-stained clinical samples where such artifacts can lead to false identification of disease.
A spectral image “cube” consists of a set of images each acquired at a different wavelength. Spectral image sets can be acquired of samples that are reflective, transmissive, or fluorescent. In many cases of interest, the final acquired image arises from the combined effect of various independent, often co-localized, spectrally-varying components present in the sample or scene being imaged. In linear spectral unmixing (LSU), it is assumed that the final acquired spectral image cube may be expressed as a linear combination of the contribution from each of these independent elements (the “reference spectra”). The mathematics of LSU allows the final image to be decomposed into separate “rule images”, one for each reference spectrum, with the pixel-by-pixel intensity of each rule image corresponding to the relative amount of that spectral component calculated to be present in the acquired image.